Preclinical validation and imaging of Wnt-induced repair in human 3D lung tissue cultures

Eur Respir J. 2015 Oct;46(4):1150-66. doi: 10.1183/09031936.00183214. Epub 2015 Apr 30.

Abstract

Chronic obstructive pulmonary disease (COPD) is characterised by a progressive loss of lung tissue. Inducing repair processes within the adult diseased lung is of major interest and Wnt/β-catenin signalling represents a promising target for lung repair. However, the translation of novel therapeutic targets from model systems into clinical use remains a major challenge.We generated murine and patient-derived three-dimensional (3D) ex vivo lung tissue cultures (LTCs), which closely mimic the 3D lung microenvironment in vivo. Using two well-known glycogen synthase kinase-3β inhibitors, lithium chloride (LiCl) and CHIR 99021 (CT), we determined Wnt/β-catenin-driven lung repair processes in high spatiotemporal resolution using quantitative PCR, Western blotting, ELISA, (immuno)histological assessment, and four-dimensional confocal live tissue imaging.Viable 3D-LTCs exhibited preserved lung structure and function for up to 5 days. We demonstrate successful Wnt/β-catenin signal activation in murine and patient-derived 3D-LTCs from COPD patients. Wnt/β-catenin signalling led to increased alveolar epithelial cell marker expression, decreased matrix metalloproteinase-12 expression, as well as altered macrophage activity and elastin remodelling. Importantly, induction of surfactant protein C significantly correlated with disease stage (per cent predicted forced expiratory volume in 1 s) in patient-derived 3D-LTCs.Patient-derived 3D-LTCs represent a valuable tool to analyse potential targets and drugs for lung repair. Enhanced Wnt/β-catenin signalling attenuated pathological features of patient-derived COPD 3D-LTCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Animals
  • Cells, Cultured
  • Disease Models, Animal
  • Emphysema / physiopathology
  • Enzyme-Linked Immunosorbent Assay
  • Epithelial Cells / cytology
  • Female
  • Glycogen Synthase Kinase 3 / antagonists & inhibitors
  • Glycogen Synthase Kinase 3 beta
  • Humans
  • Lithium Chloride / chemistry
  • Lung / cytology*
  • Lung / physiopathology
  • Macrophages, Alveolar / cytology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Confocal
  • Middle Aged
  • Pulmonary Disease, Chronic Obstructive / metabolism
  • Pulmonary Disease, Chronic Obstructive / physiopathology*
  • Pyridines / chemistry
  • Pyrimidines / chemistry
  • Signal Transduction
  • Swine
  • Wnt Proteins / metabolism*
  • Wound Healing
  • beta Catenin / metabolism

Substances

  • Chir 99021
  • Pyridines
  • Pyrimidines
  • Wnt Proteins
  • beta Catenin
  • GSK3B protein, human
  • Glycogen Synthase Kinase 3 beta
  • Gsk3b protein, mouse
  • Glycogen Synthase Kinase 3
  • Lithium Chloride